Balamurugan A N, Ramakrishna Banumathi, Gunasekaran S
Departments of Physiology and Pathology, Christian Medical College and Hospital, Vellore 632 002, TN, India.
Diabetes Res Clin Pract. 2004 Oct;66(1):13-21. doi: 10.1016/j.diabres.2004.02.012.
We describe a simple stationary digestion method of islet isolation and separation by various density gradients from monkey pancreas (Macaca radiata radiata). Effective method, different types and concentrations of collagenase were standardized. Sigma type XI collagenase yielded >1000 islets/gram pancreas at the concentration of 4 mg/ml and 3 ml Hank's/gram pancreas. Slow digestion with less concentration of collagenase was suitable for monkey islet isolation. Discontinuous density gradients of bovine serum albumin (BSA) and dextran were compared with standard Ficoll for separation of islets. Islet yield (1038 +/- 81), insulin secretory response (stimulation index, S.I.11) and histological examination revealed dextran gradients were more appropriate for monkey islets when compared to BSA and Ficoll. Insulin secretory characteristics of monkey islets were studied by exposing them to low and high concentrations glucose (S.I.11.5), arginine (S.I.4.2), leucine (S.I.2.3) and tolbutamide (S.I.1.7). The results indicated that the magnitude of glucose induced insulin secretion of monkey islet is about half as that of rat and mouse islets. However, it is higher than that of porcine and bovine islets. In conclusion, the knowledge of insulin secretory ability of Indian bonnet monkey islets together with the techniques of isolation and separation are useful tool for diabetic research especially islet transplantation.
我们描述了一种简单的静态消化方法，用于从恒河猴（Macaca radiata radiata）胰腺中通过各种密度梯度分离胰岛。对有效的方法进行了标准化，确定了不同类型和浓度的胶原酶。Sigma XI型胶原酶在浓度为4 mg/ml且每克胰腺使用3 ml Hank's液时，每克胰腺可产生>1000个胰岛。用较低浓度的胶原酶进行缓慢消化适合于恒河猴胰岛的分离。将牛血清白蛋白（BSA）和葡聚糖的不连续密度梯度与标准Ficoll用于胰岛分离进行了比较。胰岛产量（1038±81）、胰岛素分泌反应（刺激指数，S.I.11）和组织学检查显示，与BSA和Ficoll相比，葡聚糖梯度更适合于恒河猴胰岛。通过将恒河猴胰岛暴露于低浓度和高浓度葡萄糖（S.I.11.5）、精氨酸（S.I.4.2）、亮氨酸（S.I.2.3）和甲苯磺丁脲（S.I.1.7）来研究其胰岛素分泌特性。结果表明，葡萄糖诱导的恒河猴胰岛胰岛素分泌量约为大鼠和小鼠胰岛的一半。然而，它高于猪和牛胰岛。总之，印度冠叶猴胰岛胰岛素分泌能力的知识以及分离和纯化技术是糖尿病研究尤其是胰岛移植的有用工具。
